Zika virus (ZIKV), a flavivirus, has recently spread to the Americas and is currently associated with a major pandemic in human populations in South, Central, and North America. ZIKV infections in pregnant women has been associated with severe birth defects, notably microcephaly; thus, it is critical to understand the epidemiology and spread of this viral infection. ZIKV, which can be spread by mosquitos, was first isolated in Africa from non-human primates, suggesting that these animals may serve as a reservoir for the virus. However, much less is known about the reservoirs or spread of the virus in the Americas. Countries where outbreaks are occurring, including Brazil, Columbia, Ecuador and Mexico, are home to a number of species of Neotropical primates, and the human and non-human primate populations occupy overlapping areas. This suggests that there is the potential for these animals to be a reservoir for maintenance and spread of the virus in human populations. Our first aim is to determine whether Neotropical primates are a potential reservoir for ZIKV. We have assembled an expert team of primatologists, virologists, enteric infectious disease researchers, and epidemiologist who will work collaboratively to answer this question. We will sample 12 species of Neotropical primates at 8 field sites in 4 countries. We will determine the presence of active infection or carriage of ZIKV by quantitative RT-PCR, and we will test for antibodies against the virus as an indicator of prior exposure. Additionally we will obtain blood samples from human volunteers in proximal geographic areas to determine if human infection positively correlates with ZIKV presence in Neotropical primates. Molecular phylogenetic analysis of ZIKV gene sequences from human and non-human primates will help determine the epidemiology and spread of the virus in these populations. Because obtaining blood samples from the animals is a difficult and invasive procedure, our second aim is to develop a rapid, noninvasive screen for the virus. Some flaviviruses are shed in the feces of primates, therefore we will analyze fecal samples from primates that have ZIKV-positive blood cultures or serology to determine whether the virus is present. Urine, which has been used to detect ZIKV, will be an alternative method. We will modify the protocol to optimize ZIKV detection from this non-invasive sampling method. The results of this study will provide critical information on the role of Neotropical primates in the spread of Zika in the Americas.